Sea island cotton receptor analogous protein kinase gene and its application
A technology of protein kinase gene and receptor, which is applied in the field of cloning of receptor-like protein kinase gene of sea island cotton, to achieve considerable economic benefits, reduce losses, and improve the effect of verticillium wilt resistance
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[0063] 1. Design degenerate primers using the conserved domains of kinase-like disease-resistant genes that have been cloned in other crops, and perform PCR amplification from Haidao Mianhai 7124. Fragments of expected size were recovered, and the recovered fragments were ligated into pGEM-T easy Vector (Promega). Pick 10 positive clones and send them to Shanghai Handsome Biotechnology Co., Ltd. for sequencing.
[0064] 2. Sequencing results After manual removal of vector sequences by DNAstar software, BLASTx analysis was performed in the non-redundant protein database of NCBI.
[0065] 3. The 7 sequences related to kinases in the BLASTx analysis results were analyzed with DNAstar for continuous open reading frames (ORFs), and 3 sequences with continuous ORFs were found (sequence numbers: L354, L355, L361, respectively).
[0066] 4. Design specific primers using 3 sequences with continuous ORFs. Analysis of its expression in different periods in root tissues of pirate cotton...
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